Saltini C, Hance A J, Ferrans V J, Basset F, Bitterman P B, Crystal R G
Am Rev Respir Dis. 1984 Oct;130(4):650-8. doi: 10.1164/arrd.1984.130.4.650.
Quantification of the differential cell count and total number of cells recovered from the lower respiratory tract by bronchoalveolar lavage is a valuable technique for evaluating the alveolitis of patients with inflammatory disorders of the lower respiratory tract. The most commonly used technique for the evaluation of cells recovered by lavage has been to concentrate cells by centrifugation and then to determine total cell number using a hemocytometer and differential cell count from a Wright-Glemsa-stained cytocentrifuge preparation. However, we have noted that the percentage of small cells present in the original cell suspension recovered by lavage is greater than the percentage of lymphocytes identified on cytocentrifuge preparations. Therefore, we developed procedures for determining differential cell counts on lavage cells collected on Millipore filters and stained with hematoxylin-eosin (filter preparations) and compared the results of differential cell counts performed on filter preparations with those obtained using cytocentrifuge preparations. When cells recovered by lavage were collected on filter preparations, accurate differential cell counts were obtained, as confirmed by performing differential cell counts on cell mixtures of known composition, and by comparing differential cell counts obtained using filter preparations stained with hematoxylin-eosin with those obtained using filter preparations stained with a peroxidase cytochemical stain. The morphology of cells displayed on filter preparations was excellent, and interobserver variability in quantitating cell types recovered by lavage was less than 3%.(ABSTRACT TRUNCATED AT 250 WORDS)
通过支气管肺泡灌洗对下呼吸道回收的细胞进行分类计数和细胞总数定量,是评估下呼吸道炎症性疾病患者肺泡炎的一项有价值的技术。评估灌洗回收细胞最常用的技术是通过离心浓缩细胞,然后使用血细胞计数器确定细胞总数,并从经瑞氏-吉姆萨染色的细胞离心涂片制备物中进行分类计数。然而,我们注意到,灌洗回收的原始细胞悬液中小细胞的百分比大于细胞离心涂片制备物中鉴定出的淋巴细胞百分比。因此,我们开发了在密理博滤膜上收集并经苏木精-伊红染色(滤膜制备物)的灌洗细胞分类计数程序,并将滤膜制备物上的分类计数结果与细胞离心涂片制备物的结果进行比较。当灌洗回收的细胞收集在滤膜制备物上时,获得了准确的分类计数,这通过对已知组成的细胞混合物进行分类计数,以及将苏木精-伊红染色的滤膜制备物与过氧化物酶细胞化学染色的滤膜制备物获得的分类计数进行比较得到证实。滤膜制备物上显示的细胞形态极佳,灌洗回收细胞类型定量的观察者间差异小于3%。(摘要截短于250字)